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Evaluation of Genetic Variations in Maize Seedlings Exposed to Electric Field Based on Protein and DNA Markers.

Al-Huqail AA, Abdelhaliem E - Biomed Res Int (2015)

Bottom Line: Leucine-aminopeptidase, peroxidase, and catalase isozymes showed the highest values of polymorphism (100%) based on zymograms number, relative front (Rf ), and optical intensity while esterase isozyme generated polymorphism value of 83.33%.Amino acids were analyzed using high-performance liquid chromatography, which revealed the presence of 17 amino acids of variable contents ranging from 22.65% to 28.09%.RAPD revealed that 78 amplified DNA products had highly polymorphism value (95.08%) based on band numbers, with variable sizes ranging from 120 to 992 base pairs and band intensity.

View Article: PubMed Central - PubMed

Affiliation: Plant Physiology, Botany and Microbiology Department, Science College, King Saud University, P.O. Box 22452, Riyadh 11495, Saudi Arabia.

ABSTRACT
The current study analyzed proteins and nuclear DNA of electric fields (ELF) exposed and nonexposed maize seedlings for different exposure periods using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), isozymes, random amplified polymorphic DNA (RAPD), and comet assay, respectively. SDS-PAGE analysis revealed total of 46 polypeptides bands with different molecular weights ranging from 186.20 to 36.00 KDa. It generated distinctive polymorphism value of 84.62%. Leucine-aminopeptidase, peroxidase, and catalase isozymes showed the highest values of polymorphism (100%) based on zymograms number, relative front (Rf ), and optical intensity while esterase isozyme generated polymorphism value of 83.33%. Amino acids were analyzed using high-performance liquid chromatography, which revealed the presence of 17 amino acids of variable contents ranging from 22.65% to 28.09%. RAPD revealed that 78 amplified DNA products had highly polymorphism value (95.08%) based on band numbers, with variable sizes ranging from 120 to 992 base pairs and band intensity. Comet assay recorded the highest extent of nuclear DNA damage as percentage of tailed DNA (2.38%) and tail moment unit (5.36) at ELF exposure of maize nuclei for 5 days. The current study concluded that the longer ELF exposing periods had genotoxic stress on macromolecules of maize cells and biomarkers used should be augmented for reliable estimates of genotoxicity after exposure of economic plants to ELF stressors.

No MeSH data available.


Related in: MedlinePlus

RAPD products of DNA fragments extracted from ELF exposed and nonexposed maize seedlings using 5 random primers.
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fig3: RAPD products of DNA fragments extracted from ELF exposed and nonexposed maize seedlings using 5 random primers.

Mentions: RAPD analysis was employed in the present study to evaluate the extent of the DNA alterations in ELF exposed and nonexposed maize leaves as shown in Table 7 and Figure 3. Twenty random primers were used for the RAPD analysis, in which only five primers of them (P-02, 06, 08, 10, and 14) succeeded to produce clear reproducible DNA bands and gave satisfactory results with many alterations in the RAPD profiles. In total, Seventy-eight (78) amplified DNA products were scored; these bands varied in number of specific random sequences, band intensity, and sizes, ranging from 120 to 992 bp. An average of 15.60 bands per primer was scored. RAPD analysis generated highly polymorphism values of 95.16% and 75.64% (59 in total) DNA polymorphic bands, of which 51 bands were unique with value 65.35% and 7 bands were nonunique with value 8.97% in addition to 3 bands which were monomorphic with value 3.85%. The maximum number of gene products (23 bands) with value 29.49% was observed at ELF treatment T5, whereas the minimum number of bands (16) with value 20.51% was at ELF treatment T1 compared to zero treatment T0 at which RAPD analysis scored 21 bands with value 26.92%. On the other hand, the highest number of gene products (21 bands) was generated by primer P-14, whereas the lowest number (13 bands) was generated by the primers 02 and 06. Furthermore, the highest value of polymorphism (100%) was revealed by the primer −10 and 14, whereas the lowest one (87.50%) is revealed by primer 02.


Evaluation of Genetic Variations in Maize Seedlings Exposed to Electric Field Based on Protein and DNA Markers.

Al-Huqail AA, Abdelhaliem E - Biomed Res Int (2015)

RAPD products of DNA fragments extracted from ELF exposed and nonexposed maize seedlings using 5 random primers.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4491376&req=5

fig3: RAPD products of DNA fragments extracted from ELF exposed and nonexposed maize seedlings using 5 random primers.
Mentions: RAPD analysis was employed in the present study to evaluate the extent of the DNA alterations in ELF exposed and nonexposed maize leaves as shown in Table 7 and Figure 3. Twenty random primers were used for the RAPD analysis, in which only five primers of them (P-02, 06, 08, 10, and 14) succeeded to produce clear reproducible DNA bands and gave satisfactory results with many alterations in the RAPD profiles. In total, Seventy-eight (78) amplified DNA products were scored; these bands varied in number of specific random sequences, band intensity, and sizes, ranging from 120 to 992 bp. An average of 15.60 bands per primer was scored. RAPD analysis generated highly polymorphism values of 95.16% and 75.64% (59 in total) DNA polymorphic bands, of which 51 bands were unique with value 65.35% and 7 bands were nonunique with value 8.97% in addition to 3 bands which were monomorphic with value 3.85%. The maximum number of gene products (23 bands) with value 29.49% was observed at ELF treatment T5, whereas the minimum number of bands (16) with value 20.51% was at ELF treatment T1 compared to zero treatment T0 at which RAPD analysis scored 21 bands with value 26.92%. On the other hand, the highest number of gene products (21 bands) was generated by primer P-14, whereas the lowest number (13 bands) was generated by the primers 02 and 06. Furthermore, the highest value of polymorphism (100%) was revealed by the primer −10 and 14, whereas the lowest one (87.50%) is revealed by primer 02.

Bottom Line: Leucine-aminopeptidase, peroxidase, and catalase isozymes showed the highest values of polymorphism (100%) based on zymograms number, relative front (Rf ), and optical intensity while esterase isozyme generated polymorphism value of 83.33%.Amino acids were analyzed using high-performance liquid chromatography, which revealed the presence of 17 amino acids of variable contents ranging from 22.65% to 28.09%.RAPD revealed that 78 amplified DNA products had highly polymorphism value (95.08%) based on band numbers, with variable sizes ranging from 120 to 992 base pairs and band intensity.

View Article: PubMed Central - PubMed

Affiliation: Plant Physiology, Botany and Microbiology Department, Science College, King Saud University, P.O. Box 22452, Riyadh 11495, Saudi Arabia.

ABSTRACT
The current study analyzed proteins and nuclear DNA of electric fields (ELF) exposed and nonexposed maize seedlings for different exposure periods using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), isozymes, random amplified polymorphic DNA (RAPD), and comet assay, respectively. SDS-PAGE analysis revealed total of 46 polypeptides bands with different molecular weights ranging from 186.20 to 36.00 KDa. It generated distinctive polymorphism value of 84.62%. Leucine-aminopeptidase, peroxidase, and catalase isozymes showed the highest values of polymorphism (100%) based on zymograms number, relative front (Rf ), and optical intensity while esterase isozyme generated polymorphism value of 83.33%. Amino acids were analyzed using high-performance liquid chromatography, which revealed the presence of 17 amino acids of variable contents ranging from 22.65% to 28.09%. RAPD revealed that 78 amplified DNA products had highly polymorphism value (95.08%) based on band numbers, with variable sizes ranging from 120 to 992 base pairs and band intensity. Comet assay recorded the highest extent of nuclear DNA damage as percentage of tailed DNA (2.38%) and tail moment unit (5.36) at ELF exposure of maize nuclei for 5 days. The current study concluded that the longer ELF exposing periods had genotoxic stress on macromolecules of maize cells and biomarkers used should be augmented for reliable estimates of genotoxicity after exposure of economic plants to ELF stressors.

No MeSH data available.


Related in: MedlinePlus